Development and characterization of fluorescent pH sensors based on porous silica and hydrogel support matrices

The hydrogen ion activity (pH) is a very important parameter in environment monitoring, biomedical research and other applications. Optical pH sensors have several advantages over traditional potentiometric pH measurement, such as high sensitivity, no need of constant calibration, easy for miniaturization and possibility for remote sensing. Several pH indicators has been successfully immobilized in three different solid porous materials to use as pH sensing probes. The fluorescent pH indicator fluorescein-5-isothiocyanate (FITC) was covalently bound onto the internal surface of porous silica (pore size ~10 nm) and retained its pH sensitivity. The excited state pK* a of FITC in porous silica (5.58) was slightly smaller than in solution (5.68) due to the free silanol groups (Si-OH) on the silica surface. The pH sensitive range for this probe is pH 4.5 - 7.0 with an error less than 0.1 pH units. The probe response was reproducible and stable for at least four month, stored in DI water, but exhibit a long equilibrium of up to 100 minutes. Sol-gel based pH sensors were developed with immobilization of two fluorescent pH indicators fluorescein-5-(and-6)-sulfonic acid, trisodium salt (FS) and 8-hydroxypyrene- 1,3,6-trisulfonic acid (HPTS) through physical entrapment. Prior to immobilization, the indicators were ion-paired with a common surfactant hexadecyltrimethylammonium bromide (CTAB) in order to prevent leaching. The sol-gel films were synthesized through the hydrolysis of two different precursors, ethyltriethoxysilane (ETEOS) and 3- glycidoxypropyltrimethoxysilane (GPTMS) and deposited on a quartz slide through spin coating. The pK a of the indicators immobilized in sol-gel films was much smaller than in solutions due to silanol groups on the inner surface of the sol-gel films and ammonium groups from the surrounding surfactants. Unlike in solution, the apparent pK a of the indicators in sol-gel films increased with increasing ionic strength. The equilibrium time for these sensors was within 5 minutes (with film thickness of ~470 nm). Polyethylene glycol (PEG) hydrogel was of interest for optical pH sensor development because it is highly proton permeable, transparent and easy to synthesize. pH indicators can be immobilized in hydrogel through physical entrapment and copolymerization. FS and HPTS ion-pairs were physically entrapped in hydrogel matrix synthesized via free radical initiation. For covalent immobilization, three indicators, 6,8-dihydroxypyrene-1,3- disulfonic acid (DHPDS), 2,7-dihydroxynaphthalene-3,6-disulfonic acid (DHNDS) and cresol red were first reacted with methacrylic anhydride (MA) to form methacryloylanalogs for copolymerization. These hydrogels were synthesized in aqueous solution with a redox initiation system. The thickness of the hydrogel film is controlled as ~ 0.5 cm and the porosity can be adjusted with the percentage of polyethylene glycol in the precursor solutions. The pK a of the indicators immobilized in the hydrogel both physically and covalently were higher than in solution due to the medium effect. The sensors are stable and reproducible with a short equilibrium time (less than 4 minutes). In addition, the color change of cresol red immobilized hydrogel is vivid from yellow (acidic condition) to purple (basic condition). Due to covalently binding, cresol red was not leaching out from the hydrogel, making it a good candidate of reusable "pH paper".

Iowa State Daily (January 9, 2012)

ContentsLaunching his legacyOpen house unveils renovated State GymFirst-time voters impact results of Iowa caucusesFriends, teammates remember AndaryDwolla COO earns award for innovationTaking action, giving voice at caucuses

A Pt(II) complex with both a phenanthroline and a tetrathiafulvalene-extended dithiolate ligand: Synthesis, crystal structure, electrochemical and spectroscopic properties

The reaction of 4,5-bis(2'-cyanoethylsulfanyl)-4',5'-dipropylthiotetrathiafulvalene with Pt(phen)Cl-2 (phen = 1,10-phenanthroline) with CsOH as base in CH3OH-THE affords the target complex I in 44% yield. This complex crystallizes in the monoclinic space group P2(1)/c, M = 790.01, a = 12.1732(12), b = 15.851(2), c = 14.5371(16) angstrom, beta = 107.693(12)degrees, V = 2672.4(5) angstrom(3) and Z = 4. It undergoes two reversible single-electron oxidation and two irreversible reduction processes. An intense electronic absorption band at 15200 cm(-1) (658 nm) in CH2Cl2 is assigned to the intramolecular mixed metal/ligand-to-ligand charge transfer (LLCT) from a tetrathiafulvalene-extended dithiolate-based HOMO to a phenanthroline-based LUMO. This band shifts hypsochromically with increasing solvent polarity. Systematic changes in the optical spectra upon oxidation allow precise tuning of the oxidation states of 1 and reversible control over its optical properties. Irradiation of 1 at 15625 cm(-1) (640 nm) in glassy solution below 150K results in emission from the (LLCT)-L-3 excited state. GRAPHICS (C) 2013 Elsevier Ltd. All rights reserved.

Tetrathiafulvalene - Benzothiadiazoles as Redox-Tunable Donor - Acceptor Systems: Synthesis and Photophysical Study

Electrochemical and photophysical analysis of new donoracceptor systems 2 and 3, in which a benzothiadiazole (BTD) unit is covalently linked to a tetrathiafulvalene (TTF) core, have verified that the lowest excited state can be ascribed to an intramolecular-charge-transfer (ICT) (TTF)*(benzothiadiazole) transition. Owing to better overlap of the HOMO and LUMO in the fused scaffold of compound 3, the intensity of the 1ICT band is substantially higher compared to that in compound 2. The corresponding CT fluorescence is also observed in both cases. The radical cation TTF+. is easily observed through chemical and electrochemical oxidation by performing steady-state absorption experiments. Interestingly, compound 2 is photo-oxidized under aerobic conditions.

Optical and scintillation properties of CsBa2I5:Eu2+

The scintillation and luminescence properties of pure CsBa2I5 and CsBa2I5 doped with 0.5% Eu and 5% Eu were studied between 78 K and 600 K. Single crystals were grown by the vertical Bridgman method from the melt. CsBa2I5:5% Eu showed a light yield of 80,000 photons/MeV, an energy resolution of 2.3% for the 662 key full absorption peak, and an excellent proportional response. Two broad emission bands centered at 400 nm and 600 nm were observed in the radioluminescence spectrum of pure CsBa2I5. The Eu2+ 5d-4f emission band was observed at 430 nm. The radiative lifetime of the Eu2+ excited state was determined as 350 ns. With increasing temperature and Eu concentration the Eu2+ emission shifts to longer wavelengths and its decay time lengthens as a result of self-absorption of the Eu2+ emission. Multiple thermoluminescence glow peaks and a sharp decrease of the light yield at temperatures below 200 K were observed and related to the presence of the charge carrier traps in CsBa2I5:Eu.

Energy and charge transfer dynamics in DNA based light harvesting antennae

DNA can serve as a versatile scaffold for chromophore assemblies. For example, light-harvesting antennae have been realized by incorporating phenanthrene and pyrene building blocks into DNA strands. It was shown that by exciting at 320 nm (absorption of phenanthrene), an emission at 450 nm is observed which corresponds to a phenanthrene-pyrene exciplex. The more phenanthrenes are added into the DNA duplex, the higher is the fluorescence intensity with no significant change in quantum yield. This shows that phenanthrene acts as a donor and efficiently transfers the excitation energy to the pyrene. Up to now, the mechanism of this energy transfer and exciplex formation is not known. Therefore, we first aim at studying the photo-cycle of such DNA assemblies through transient absorption spectroscopy. Based on the results, we will explore ways to manipulate the energy transfer by application of intense THz fields. Ground as well as excited state Stark effect dynamics will be investigated.

Experimental and Calculated Spectra of π-Stacked Mild Charge-Transfer Complexes: Jet-Cooled Perylene·(Tetrachloroethene) n , n = 1,2

The S0 ↔ S1 spectra of the mild charge-transfer (CT) complexes perylene·tetrachloroethene (P·4ClE) and perylene·(tetrachloroethene)2 (P·(4ClE)2) are investigated by two-color resonant two-photon ionization (2C-R2PI) and dispersed fluorescence spectroscopy in supersonic jets. The S0 → S1 vibrationless transitions of P·4ClE and P·(4ClE)2 are shifted by δν = −451 and −858 cm–1 relative to perylene, translating to excited-state dissociation energy increases of 5.4 and 10.3 kJ/mol, respectively. The red shift is ∼30% larger than that of perylene·trans-1,2-dichloroethene; therefore, the increase in chlorination increases the excited-state stabilization and CT character of the interaction, but the electronic excitation remains largely confined to the perylene moiety. The 2C-R2PI and fluorescence spectra of P·4ClE exhibit strong progressions in the perylene intramolecular twist (1au) vibration (42 cm–1 in S0 and 55 cm–1 in S1), signaling that perylene deforms along its twist coordinate upon electronic excitation. The intermolecular stretching (Tz) and internal rotation (Rc) vibrations are weak; therefore, the P·4ClE intermolecular potential energy surface (IPES) changes little during the S0 ↔ S1 transition. The minimum-energy structures and inter- and intramolecular vibrational frequencies of P·4ClE and P·(4ClE)2 are calculated with the dispersion-corrected density functional theory (DFT) methods B97-D3, ωB97X-D, M06, and M06-2X and the spin-consistent-scaled (SCS) variant of the approximate second-order coupled-cluster method, SCS-CC2. All methods predict the global minima to be π-stacked centered coplanar structures with the long axis of tetrachloroethene rotated by τ ≈ 60° relative to the perylene long axis. The calculated binding energies are in the range of −D0 = 28–35 kJ/mol. A second minimum is predicted with τ ≈ 25°, with ∼1 kJ/mol smaller binding energy. Although both monomers are achiral, both the P·4ClE and P·(4ClE)2 complexes are chiral. The best agreement for adiabatic excitation energies and vibrational frequencies is observed for the ωB97X-D and M06-2X DFT methods.

Modeling the Histidine–Phenylalanine Interaction: The NH···π Hydrogen Bond of Imidazole·Benzene

NH···π hydrogen bonds occur frequently between the amino acid side groups in proteins and peptides. Data-mining studies of protein crystals find that ~80% of the T-shaped histidine···aromatic contacts are CH···π, and only ~20% are NH···π interactions. We investigated the infrared (IR) and ultraviolet (UV) spectra of the supersonic-jet-cooled imidazole·benzene (Im·Bz) complex as a model for the NH···π interaction between histidine and phenylalanine. Ground- and excited-state dispersion-corrected density functional calculations and correlated methods (SCS-MP2 and SCS-CC2) predict that Im·Bz has a Cs-symmetric T-shaped minimum-energy structure with an NH···π hydrogen bond to the Bz ring; the NH bond is tilted 12° away from the Bz C₆ axis. IR depletion spectra support the T-shaped geometry: The NH stretch vibrational fundamental is red shifted by −73 cm⁻¹ relative to that of bare imidazole at 3518 cm⁻¹, indicating a moderately strong NH···π interaction. While the Sₒ(A1g) → S₁(B₂u) origin of benzene at 38 086 cm⁻¹ is forbidden in the gas phase, Im·Bz exhibits a moderately intense Sₒ → S₁ origin, which appears via the D₆h → Cs symmetry lowering of Bz by its interaction with imidazole. The NH···π ground-state hydrogen bond is strong, De=22.7 kJ/mol (1899 cm⁻¹). The combination of gas-phase UV and IR spectra confirms the theoretical predictions that the optimum Im·Bz geometry is T shaped and NH···π hydrogen bonded. We find no experimental evidence for a CH···π hydrogen-bonded ground-state isomer of Im·Bz. The optimum NH···π geometry of the Im·Bz complex is very different from the majority of the histidine·aromatic contact geometries found in protein database analyses, implying that the CH···π contacts observed in these searches do not arise from favorable binding interactions but merely from protein side-chain folding and crystal-packing constraints. The UV and IR spectra of the imidazole·(benzene)₂ cluster are observed via fragmentation into the Im·Bz+ mass channel. The spectra of Im·Bz and Im·Bz₂ are cleanly separable by IR hole burning. The UV spectrum of Im·Bz₂ exhibits two 000 bands corresponding to the Sₒ → S₁ excitations of the two inequivalent benzenes, which are symmetrically shifted by −86/+88 cm⁻¹ relative to the 000 band of benzene.

Molecular control of cardiac sodium homeostasis in health and disease.

INTRODUCTION Cardiac myocytes utilize three high-capacity Na transport processes whose precise function can determine myocyte fate and the triggering of arrhythmias in pathological settings. We present recent results on the regulation of all three transporters that may be important for an understanding of cardiac function during ischemia/reperfusion episodes. METHODS AND RESULTS Refined ion selective electrode (ISE) techniques and giant patch methods were used to analyze the function of cardiac Na/K pumps, Na/Ca exchange (NCX1), and Na/H exchange (NHE1) in excised cardiac patches and intact myocytes. To consider results cohesively, simulations were developed that account for electroneutrality of the cytoplasm, ion homeostasis, water homeostasis (i.e., cell volume), and cytoplasmic pH. The Na/K pump determines the average life-time of Na ions (3-10 minutes) as well as K ions (>30 minutes) in the cytoplasm. The long time course of K homeostasis can determine the time course of myocyte volume changes after ion homeostasis is perturbed. In excised patches, cardiac Na/K pumps turn on slowly (-30 seconds) with millimolar ATP dependence, when activated for the first time. In steady state, however, pumps are fully active with <0.2 mM ATP and are nearly unaffected by high ADP (2 mM) and Pi (10 mM) concentrations as may occur in ischemia. NCX1s appear to operate with slippage that contributes to background Na influx and inward current in heart. Thus, myocyte Na levels may be regulated by the inactivation reactions of the exchanger which are both Na- and proton-dependent. NHE1 also undergo strong Na-dependent inactivation, whereby a brief rise of cytoplasmic Na can cause inactivation that persists for many minutes after cytoplasmic Na is removed. This mechanism is blocked by pertussis toxin, suggesting involvement of a Na-dependent G-protein. Given that maximal NCX1- and NHE1-mediated ion fluxes are much greater than maximal Na/K pump-mediated Na extrusion in myocytes, the Na-dependent inactivation mechanisms of NCX1 and NHE1 may be important determinants of cardiac Na homeostasis. CONCLUSIONS Na/K pumps appear to be optimized to continue operation when energy reserves are compromised. Both NCX1 and NHE1 activities are regulated by accumulation of cytoplasmic Na. These principles may importantly control cardiac cytoplasmic Na and promote myocyte survival during ischemia/reperfusion episodes by preventing Ca overload.

Output Impedance Correction Circuit (OICC): A New Concept to Improve the Dynamic Response of DC/DC Converters with Additional Energy Path

El desarrollo da las nuevas tecnologías permite a los ingenieros llevar al límite el funcionamiento de los circuitos integrados (Integrated Circuits, IC). Las nuevas generaciones de procesadores, DSPs o FPGAs son capaces de procesar la información a una alta velocidad, con un alto consumo de energía, o esperar en modo de baja potencia con el mínimo consumo posible. Esta gran variación en el consumo de potencia y el corto tiempo necesario para cambiar de un nivel al otro, afecta a las especificaciones del Módulo de Regulador de Tensión (Voltage Regulated Module, VRM) que alimenta al IC. Además, las características adicionales obligatorias, tales como adaptación del nivel de tensión (Adaptive Voltage Positioning, AVP) y escalado dinámico de la tensión (Dynamic Voltage Scaling, DVS), imponen requisitos opuestas en el diseño de la etapa de potencia del VRM. Para poder soportar las altas variaciones de los escalones de carga, el condensador de filtro de salida del VRM se ha de sobredimensionar, penalizando la densidad de energía y el rendimiento durante la operación de DVS. Por tanto, las actuales tendencias de investigación se centran en mejorar la respuesta dinámica del VRM, mientras se reduce el tamaño del condensador de salida. La reducción del condensador de salida lleva a menor coste y una prolongación de la vida del sistema ya que se podría evitar el uso de condensadores voluminosos, normalmente implementados con condensadores OSCON. Una ventaja adicional es que reduciendo el condensador de salida, el DVS se puede realizar más rápido y con menor estrés de la etapa de potencia, ya que la cantidad de carga necesaria para cambiar la tensión de salida es menor. El comportamiento dinámico del sistema con un control lineal (Control Modo Tensión, VMC, o Control Corriente de Pico, Peak Current Mode Control, PCMC,…) está limitado por la frecuencia de conmutación del convertidor y por el tamaño del filtro de salida. La reducción del condensador de salida se puede lograr incrementando la frecuencia de conmutación, así como incrementando el ancho de banda del sistema, y/o aplicando controles avanzados no-lineales. Usando esos controles, las variables del estado se saturan para conseguir el nuevo régimen permanente en un tiempo mínimo, así como el filtro de salida, más específicamente la pendiente de la corriente de la bobina, define la respuesta de la tensión de salida. Por tanto, reduciendo la inductancia de la bobina de salida, la corriente de bobina llega más rápido al nuevo régimen permanente, por lo que una menor cantidad de carga es tomada del condensador de salida durante el tránsito. El inconveniente de esa propuesta es que el rendimiento del sistema es penalizado debido al incremento de pérdidas de conmutación y las corrientes RMS. Para conseguir tanto la reducción del condensador de salida como el alto rendimiento del sistema, mientras se satisfacen las estrictas especificaciones dinámicas, un convertidor multifase es adoptado como estándar para aplicaciones VRM. Para asegurar el reparto de las corrientes entre fases, el convertidor multifase se suele implementar con control de modo de corriente. Para superar la limitación impuesta por el filtro de salida, la segunda posibilidad para reducir el condensador de salida es aplicar alguna modificación topológica (Topologic modifications) de la etapa básica de potencia para incrementar la pendiente de la corriente de bobina y así reducir la duración de tránsito. Como el transitorio se ha reducido, una menor cantidad de carga es tomada del condensador de salida bajo el mismo escalón de la corriente de salida, con lo cual, el condensador de salida se puede reducir para lograr la misma desviación de la tensión de salida. La tercera posibilidad para reducir el condensador de salida del convertidor es introducir un camino auxiliar de energía (additional energy path, AEP) para compensar el desequilibrio de la carga del condensador de salida reduciendo consecuentemente la duración del transitorio y la desviación de la tensión de salida. De esta manera, durante el régimen permanente, el sistema tiene un alto rendimiento debido a que el convertidor principal con bajo ancho de banda es diseñado para trabajar con una frecuencia de conmutación moderada para conseguir requisitos estáticos. Por otro lado, el comportamiento dinámico durante los transitorios es determinado por el AEP con un alto ancho de banda. El AEP puede ser implementado como un camino resistivo, como regulador lineal (Linear regulator, LR) o como un convertidor conmutado. Las dos primeras implementaciones proveen un mayor ancho de banda, acosta del incremento de pérdidas durante el transitorio. Por otro lado, la implementación del convertidor computado presenta menor ancho de banda, limitado por la frecuencia de conmutación, aunque produce menores pérdidas comparado con las dos anteriores implementaciones. Dependiendo de la aplicación, la implementación y la estrategia de control del sistema, hay una variedad de soluciones propuestas en el Estado del Arte (State-of-the-Art, SoA), teniendo diferentes propiedades donde una solución ofrece más ventajas que las otras, pero también unas desventajas. En general, un sistema con AEP ideal debería tener las siguientes propiedades: 1. El impacto del AEP a las pérdidas del sistema debería ser mínimo. A lo largo de la operación, el AEP genera pérdidas adicionales, con lo cual, en el caso ideal, el AEP debería trabajar por un pequeño intervalo de tiempo, solo durante los tránsitos; la otra opción es tener el AEP constantemente activo pero, por la compensación del rizado de la corriente de bobina, se generan pérdidas innecesarias. 2. El AEP debería ser activado inmediatamente para minimizar la desviación de la tensión de salida. Para conseguir una activación casi instantánea, el sistema puede ser informado por la carga antes del escalón o el sistema puede observar la corriente del condensador de salida, debido a que es la primera variable del estado que actúa a la perturbación de la corriente de salida. De esa manera, el AEP es activado con casi cero error de la tensión de salida, logrando una menor desviación de la tensión de salida. 3. El AEP debería ser desactivado una vez que el nuevo régimen permanente es detectado para evitar los transitorios adicionales de establecimiento. La mayoría de las soluciones de SoA estiman la duración del transitorio, que puede provocar un transitorio adicional si la estimación no se ha hecho correctamente (por ejemplo, si la corriente de bobina del convertidor principal tiene un nivel superior o inferior al necesitado, el regulador lento del convertidor principal tiene que compensar esa diferencia una vez que el AEP es desactivado). Otras soluciones de SoA observan las variables de estado, asegurando que el sistema llegue al nuevo régimen permanente, o pueden ser informadas por la carga. 4. Durante el transitorio, como mínimo un subsistema, o bien el convertidor principal o el AEP, debería operar en el lazo cerrado. Implementando un sistema en el lazo cerrado, preferiblemente el subsistema AEP por su ancho de banda elevado, se incrementa la robustez del sistema a los parásitos. Además, el AEP puede operar con cualquier tipo de corriente de carga. Las soluciones que funcionan en el lazo abierto suelen preformar el control de balance de carga con mínimo tiempo, así reducen la duración del transitorio y tienen un impacto menor a las pérdidas del sistema. Por otro lado, esas soluciones demuestran una alta sensibilidad a las tolerancias y parásitos de los componentes. 5. El AEP debería inyectar la corriente a la salida en una manera controlada, así se reduce el riesgo de unas corrientes elevadas y potencialmente peligrosas y se incrementa la robustez del sistema bajo las perturbaciones de la tensión de entrada. Ese problema suele ser relacionado con los sistemas donde el AEP es implementado como un convertidor auxiliar. El convertidor auxiliar es diseñado para una potencia baja, con lo cual, los dispositivos elegidos son de baja corriente/potencia. Si la corriente no es controlada, bajo un pico de tensión de entrada provocada por otro parte del sistema (por ejemplo, otro convertidor conectado al mismo bus), se puede llegar a un pico en la corriente auxiliar que puede causar la perturbación de tensión de salida e incluso el fallo de los dispositivos del convertidor auxiliar. Sin embargo, cuando la corriente es controlada, usando control del pico de corriente o control con histéresis, la corriente auxiliar tiene el control con prealimentación (feed-forward) de tensión de entrada y la corriente es definida y limitada. Por otro lado, si la solución utiliza el control de balance de carga, el sistema puede actuar de forma deficiente si la tensión de entrada tiene un valor diferente del nominal, provocando que el AEP inyecta/toma más/menos carga que necesitada. 6. Escalabilidad del sistema a convertidores multifase. Como ya ha sido comentado anteriormente, para las aplicaciones VRM por la corriente de carga elevada, el convertidor principal suele ser implementado como multifase para distribuir las perdidas entre las fases y bajar el estrés térmico de los dispositivos. Para asegurar el reparto de las corrientes, normalmente un control de modo corriente es usado. Las soluciones de SoA que usan VMC son limitadas a la implementación con solo una fase. Esta tesis propone un nuevo método de control del flujo de energía por el AEP y el convertidor principal. El concepto propuesto se basa en la inyección controlada de la corriente auxiliar al nodo de salida donde la amplitud de la corriente es n-1 veces mayor que la corriente del condensador de salida con las direcciones apropiadas. De esta manera, el AEP genera un condensador virtual cuya capacidad es n veces mayor que el condensador físico y reduce la impedancia de salida. Como el concepto propuesto reduce la impedancia de salida usando el AEP, el concepto es llamado Output Impedance Correction Circuit (OICC) concept. El concepto se desarrolla para un convertidor tipo reductor síncrono multifase con control modo de corriente CMC (incluyendo e implementación con una fase) y puede operar con la tensión de salida constante o con AVP. Además, el concepto es extendido a un convertidor de una fase con control modo de tensión VMC. Durante la operación, el control de tensión de salida de convertidor principal y control de corriente del subsistema OICC están siempre cerrados, incrementando la robustez a las tolerancias de componentes y a los parásitos del cirquito y permitiendo que el sistema se pueda enfrentar a cualquier tipo de la corriente de carga. Según el método de control propuesto, el sistema se puede encontrar en dos estados: durante el régimen permanente, el sistema se encuentra en el estado Idle y el subsistema OICC esta desactivado. Por otro lado, durante el transitorio, el sistema se encuentra en estado Activo y el subsistema OICC está activado para reducir la impedancia de salida. El cambio entre los estados se hace de forma autónoma: el sistema entra en el estado Activo observando la corriente de condensador de salida y vuelve al estado Idle cunado el nuevo régimen permanente es detectado, observando las variables del estado. La validación del concepto OICC es hecha aplicándolo a un convertidor tipo reductor síncrono con dos fases y de 30W cuyo condensador de salida tiene capacidad de 140μF, mientras el factor de multiplicación n es 15, generando en el estado Activo el condensador virtual de 2.1mF. El subsistema OICC es implementado como un convertidor tipo reductor síncrono con PCMC. Comparando el funcionamiento del convertidor con y sin el OICC, los resultados demuestran que se ha logrado una reducción de la desviación de tensión de salida con factor 12, tanto con funcionamiento básico como con funcionamiento AVP. Además, los resultados son comparados con un prototipo de referencia que tiene la misma etapa de potencia y un condensador de salida físico de 2.1mF. Los resultados demuestran que los dos sistemas tienen el mismo comportamiento dinámico. Más aun, se ha cuantificado el impacto en las pérdidas del sistema operando bajo una corriente de carga pulsante y bajo DVS. Se demuestra que el sistema con OICC mejora el rendimiento del sistema, considerando las pérdidas cuando el sistema trabaja con la carga pulsante y con DVS. Por lo último, el condensador de salida de sistema con OICC es mucho más pequeño que el condensador de salida del convertidor de referencia, con lo cual, por usar el concepto OICC, la densidad de energía se incrementa. En resumen, las contribuciones principales de la tesis son: • El concepto propuesto de Output Impedance Correction Circuit (OICC), • El control a nivel de sistema basado en el método usado para cambiar los estados de operación, • La implementación del subsistema OICC en lazo cerrado conjunto con la implementación del convertidor principal, • La cuantificación de las perdidas dinámicas bajo la carga pulsante y bajo la operación DVS, y • La robustez del sistema bajo la variación del condensador de salida y bajo los escalones de carga consecutiva. ABSTRACT Development of new technologies allows engineers to push the performance of the integrated circuits to its limits. New generations of processors, DSPs or FPGAs are able to process information with high speed and high consumption or to wait in low power mode with minimum possible consumption. This huge variation in power consumption and the short time needed to change from one level to another, affect the specifications of the Voltage Regulated Module (VRM) that supplies the IC. Furthermore, additional mandatory features, such as Adaptive Voltage Positioning (AVP) and Dynamic Voltage Scaling (DVS), impose opposite trends on the design of the VRM power stage. In order to cope with high load-step amplitudes, the output capacitor of the VRM power stage output filter is drastically oversized, penalizing power density and the efficiency during the DVS operation. Therefore, the ongoing research trend is directed to improve the dynamic response of the VRM while reducing the size of the output capacitor. The output capacitor reduction leads to a smaller cost and longer life-time of the system since the big bulk capacitors, usually implemented with OSCON capacitors, may not be needed to achieve the desired dynamic behavior. An additional advantage is that, by reducing the output capacitance, dynamic voltage scaling (DVS) can be performed faster and with smaller stress on the power stage, since the needed amount of charge to change the output voltage is smaller. The dynamic behavior of the system with a linear control (Voltage mode control, VMC, Peak Current Mode Control, PCMC,…) is limited by the converter switching frequency and filter size. The reduction of the output capacitor can be achieved by increasing the switching frequency of the converter, thus increasing the bandwidth of the system, and/or by applying advanced non-linear controls. Applying nonlinear control, the system variables get saturated in order to reach the new steady-state in a minimum time, thus the output filter, more specifically the output inductor current slew-rate, determines the output voltage response. Therefore, by reducing the output inductor value, the inductor current reaches faster the new steady state, so a smaller amount of charge is taken from the output capacitor during the transient. The drawback of this approach is that the system efficiency is penalized due to increased switching losses and RMS currents. In order to achieve both the output capacitor reduction and high system efficiency, while satisfying strict dynamic specifications, a Multiphase converter system is adopted as a standard for VRM applications. In order to ensure the current sharing among the phases, the multiphase converter is usually implemented with current mode control. In order to overcome the limitation imposed by the output filter, the second possibility to reduce the output capacitor is to apply Topologic modifications of the basic power stage topology in order to increase the slew-rate of the inductor current and, therefore, reduce the transient duration. Since the transient is reduced, smaller amount of charge is taken from the output capacitor under the same load current, thus, the output capacitor can be reduced to achieve the same output voltage deviation. The third possibility to reduce the output capacitor of the converter is to introduce an additional energy path (AEP) to compensate the charge unbalance of the output capacitor, consequently reducing the transient time and output voltage deviation. Doing so, during the steady-state operation the system has high efficiency because the main low-bandwidth converter is designed to operate at moderate switching frequency, to meet the static requirements, whereas the dynamic behavior during the transients is determined by the high-bandwidth auxiliary energy path. The auxiliary energy path can be implemented as a resistive path, as a Linear regulator, LR, or as a switching converter. The first two implementations provide higher bandwidth, at the expense of increasing losses during the transient. On the other hand, the switching converter implementation presents lower bandwidth, limited by the auxiliary converter switching frequency, though it produces smaller losses compared to the two previous implementations. Depending on the application, the implementation and the control strategy of the system, there is a variety of proposed solutions in the State-of-the-Art (SoA), having different features where one solution offers some advantages over the others, but also some disadvantages. In general, an ideal additional energy path system should have the following features: 1. The impact on the system losses should be minimal. During its operation, the AEP generates additional losses, thus ideally, the AEP should operate for a short period of time, only when the transient is occurring; the other option is to have the AEP constantly on, but due to the inductor current ripple compensation at the output, unnecessary losses are generated. 2. The AEP should be activated nearly instantaneously to prevent bigger output voltage deviation. To achieve near instantaneous activation, the converter system can be informed by the load prior to the load-step or the system can observe the output capacitor current, which is the first system state variable that reacts on the load current perturbation. In this manner, the AEP is turned on with near zero output voltage error, providing smaller output voltage deviation. 3. The AEP should be deactivated once the new steady state is reached to avoid additional settling transients. Most of the SoA solutions estimate duration of the transient which may cause additional transient if the estimation is not performed correctly (e.g. if the main converter inductor current has higher or lower value than needed, the slow regulator of the main converter needs to compensate the difference after the AEP is deactivated). Other SoA solutions are observing state variables, ensuring that the system reaches the new steady state or they are informed by the load. 4. During the transient, at least one subsystem, either the main converter or the AEP, should be in closed-loop. Implementing a closed loop system, preferably the AEP subsystem, due its higher bandwidth, increases the robustness under system tolerances and circuit parasitic. In addition, the AEP can operate with any type of load. The solutions that operate in open loop usually perform minimum time charge balance control, thus reducing the transient length and minimizing the impact on the losses, however they are very sensitive to tolerances and parasitics. 5. The AEP should inject current at the output in a controlled manner, thus reducing the risk of high and potentially damaging currents and increasing robustness on the input voltage deviation. This issue is mainly related to the systems where AEP is implemented as auxiliary converter. The auxiliary converter is designed for small power and, as such, the MOSFETs are rated for small power/currents. If the current is not controlled, due to the some unpredicted spike in input voltage caused by some other part of the system (e.g. different converter), it may lead to a current spike in auxiliary current which will cause the perturbation of the output voltage and even failure of the switching components of auxiliary converter. In the case when the current is controlled, using peak CMC or Hysteretic Window CMC, the auxiliary converter has inherent feed-forwarding of the input voltage in current control and the current is defined and limited. Furthermore, if the solution employs charge balance control, the system may perform poorly if the input voltage has different value than the nominal, causing that AEP injects/extracts more/less charge than needed. 6. Scalability of the system to multiphase converters. As commented previously, in VRM applications, due to the high load currents, the main converters are implemented as multiphase to redistribute losses among the modules, lowering temperature stress of the components. To ensure the current sharing, usually a Current Mode Control (CMC) is employed. The SoA solutions that are implemented with VMC are limited to a single stage implementation. This thesis proposes a novel control method of the energy flow through the AEP and the main converter system. The proposed concept relays on a controlled injection of the auxiliary current at the output node where the instantaneous current value is n-1 times bigger than the output capacitor current with appropriate directions. Doing so, the AEP creates an equivalent n times bigger virtual capacitor at the output, thus reducing the output impedance. Due to the fact that the proposed concept reduces the output impedance using the AEP, it has been named the Output Impedance Correction Circuit (OICC) concept. The concept is developed for a multiphase CMC synchronous buck converter (including a single phase implementation), operating with a constant output voltage and with AVP feature. Further, it is extended to a single phase VMC synchronous buck converter. During the operation, the main converter voltage loop and the OICC subsystem capacitor current loop is constantly closed, increasing the robustness under system tolerances and circuit parasitic and allowing the system to operate with any load-current shape or pattern. According to the proposed control method, the system operates in two states: during the steady-state the system is in the Idle state and the OICC subsystem is deactivated, while during the load-step transient the system is in the Active state and the OICC subsystem is activated in order to reduce the output impedance. The state changes are performed autonomously: the system enters in the Active state by observing the output capacitor current and it returns back to the Idle state when the steady-state operation is detected by observing the state variables. The validation of the OICC concept has been done by applying it to a 30W two phase synchronous buck converter with 140μF output capacitor and with the multiplication factor n equal to 15, generating during the Active state equivalent output capacitor of 2.1mF. The OICC subsystem is implemented as single phase PCMC synchronous buck converter. Comparing the converter operation with and without the OICC the results demonstrate that the 12 times reduction of the output voltage deviation is achieved, for both basic operation and for the AVP operation. Furthermore, the results have been compared to a reference prototype which has the same power stage and a fiscal output capacitor of 2.1mF. The results show that the two systems have the same dynamic behavior. Moreover, an impact on the system losses under the pulsating load and DVS operation has been quantified and it has been demonstrated that the OICC system has improved the system efficiency, considering the losses when the system operates with the pulsating load and the DVS operation. Lastly, the output capacitor of the OICC system is much smaller than the reference design output capacitor, therefore, by applying the OICC concept the power density can be increased. In summary, the main contributions of the thesis are: • The proposed Output Impedance Correction Circuit (OICC) concept, • The system level control based on the used approach to change the states of operation, • The OICC subsystem closed-loop implementation, together with the main converter implementation, • The dynamic losses under the pulsating load and the DVS operation quantification, and • The system robustness on the capacitor impedance variation and consecutive load-steps.

Sistemas de gestión de puentes : optimización de estrategias de mantenimiento : implementación en redes locales de carreteras

En la actualidad, y en consonancia con la tendencia de “sostenibilidad” extendida a todos los campos y parcelas de la ciencia, nos encontramos con un área de estudio basado en la problemática del inevitable deterioro de las estructuras existentes, y la gestión de las acciones a realizar para mantener las condiciones de servicio de los puentes y prolongar su vida útil. Tal y como se comienza a ver en las inversiones en los países avanzados, con una larga tradición en el desarrollo de sus infraestructuras, se muestra claramente el nuevo marco al que nos dirigimos. Las nuevas tendencias van encaminadas cada vez más a la conservación y mantenimiento, reduciéndose las partidas presupuestarias destinadas a nuevas actuaciones, debido a la completa vertebración territorial que se ha ido instaurando en estos países, entre los que España se encuentra. Este nutrido patrimonio de infraestructuras viarias, que cuentan a su vez con un importante número de estructuras, hacen necesarias las labores de gestión y mantenimiento de los puentes integrantes en las mismas. Bajo estas premisas, la tesis aborda el estado de desarrollo de la implementación de los sistemas de gestión de puentes, las tendencias actuales e identificación de campos por desarrollar, así como la aplicación específica a redes de carreteras de escasos recursos, más allá de la Red Estatal. Además de analizar las diversas metodologías de formación de inventarios, realización de inspecciones y evaluación del estado de puentes, se ha enfocado, como principal objetivo, el desarrollo de un sistema específico de predicción del deterioro y ayuda a la toma de decisiones. Este sistema, adicionalmente a la configuración tradicional de criterios de formación de bases de datos de estructuras e inspecciones, plantea, de forma justificada, la clasificación relativa al conjunto de la red gestionada, según su estado de condición. Eso permite, mediante técnicas de optimización, la correcta toma de decisiones a los técnicos encargados de la gestión de la red. Dentro de los diversos métodos de evaluación de la predicción de evolución del deterioro de cada puente, se plantea la utilización de un método bilineal simplificado envolvente del ajuste empírico realizado y de los modelos markovianos como la solución más efectiva para abordar el análisis de la predicción de la propagación del daño. Todo ello explotando la campaña experimenta realizada que, a partir de una serie de “fotografías técnicas” del estado de la red de puentes gestionados obtenidas mediante las inspecciones realizadas, es capaz de mejorar el proceso habitual de toma de decisiones. Toda la base teórica reflejada en el documento, se ve complementada mediante la implementación de un Sistema de Gestión de Puentes (SGP) específico, adaptado según las necesidades y limitaciones de la administración a la que se ha aplicado, en concreto, la Dirección General de Carreteras de la Junta de Comunidades de Castilla-La Mancha, para una muestra representativa del conjunto de puentes de la red de la provincia de Albacete, partiendo de una situación en la que no existe, actualmente, un sistema formal de gestión de puentes. Tras un meditado análisis del estado del arte dentro de los Capítulos 2 y 3, se plantea un modelo de predicción del deterioro dentro del Capítulo 4 “Modelo de Predicción del Deterioro”. De la misma manera, para la resolución del problema de optimización, se justifica la utilización de un novedoso sistema de optimización secuencial elegido dentro del Capítulo 5, los “Algoritmos Evolutivos”, en sus diferentes variantes, como la herramienta matemática más correcta para distribuir adecuadamente los recursos económicos dedicados a mantenimiento y conservación de los que esta administración pueda disponer en sus partidas de presupuesto a medio plazo. En el Capítulo 6, y en diversos Anexos al presente documento, se muestran los datos y resultados obtenidos de la aplicación específica desarrollada para la red local analizada, utilizando el modelo de deterioro y optimización secuencial, que garantiza la correcta asignación de los escasos recursos de los que disponen las redes autonómicas en España. Se plantea con especial interés la implantación de estos sistemas en la red secundaria española, debido a que reciben en los últimos tiempos una mayor responsabilidad de gestión, con recursos cada vez más limitados. Finalmente, en el Capítulo 7, se plantean una serie de conclusiones que nos hacen reflexionar de la necesidad de comenzar a pasar, en materia de gestión de infraestructuras, de los estudios teóricos y los congresos, hacia la aplicación y la práctica, con un planteamiento que nos debe llevar a cambios importantes en la forma de concebir la labor del ingeniero y las enseñanzas que se imparten en las escuelas. También se enumeran las aportaciones originales que plantea el documento frente al actual estado del arte. Se plantean, de la misma manera, las líneas de investigación en materia de Sistemas de Gestión de Puentes que pueden ayudar a refinar y mejorar los actuales sistemas utilizados. In line with the development of "sustainability" extended to all fields of science, we are faced with the inevitable and ongoing deterioration of existing structures, leading nowadays to the necessary management of maintaining the service conditions and life time extension of bridges. As per the increased amounts of money that can be observed being spent in the countries with an extensive and strong tradition in the development of their infrastructure, the trend can be clearly recognized. The new tendencies turn more and more towards conservation and maintenance, reducing programmed expenses for new construction activities, in line with the already wellestablished territorial structures, as is the case for Spain. This significant heritage of established road infrastructure, consequently containing a vast number of structures, imminently lead to necessary management and maintenance of the including bridges. Under these conditions, this thesis focusses on the status of the development of the management implementation for bridges, current trends, and identifying areas for further development. This also includes the specific application to road networks with limited resources, beyond the national highways. In addition to analyzing the various training methodologies, inventory inspections and condition assessments of bridges, the main objective has been the development of a specific methodology. This methodology, in addition to the traditional system of structure and inspection database training criteria, sustains the classification for the entire road network, according to their condition. This allows, through optimization techniques, for the correct decision making by the technical managers of the network. Among the various methods for assessing the evolution forecast of deterioration of each bridge, a simplified bilinear envelope adjustment made empirical method and Markov models as the most effective solution to address the analysis of predicting the spread of damage, arising from a "technical snapshot" obtained through inspections of the condition of the bridges included in the investigated network. All theoretical basis reflected in the document, is completed by implementing a specific Bridges Management System (BMS), adapted according to the needs and limitations of the authorities for which it has been applied, being in this case particularly the General Highways Directorate of the autonomous region of Castilla-La Mancha, for a representative sample of all bridges in the network in the province of Albacete, starting from a situation where there is currently no formal bridge management system. After an analysis of the state of the art in Chapters 2 and 3, a new deterioration prediction model is developed in Chapter 4, "Deterioration Prediction Model". In the same way, to solve the optimization problem is proposed the use of a singular system of sequential optimization elected under Chapter 5, the "Evolutionary Algorithms", the most suitable mathematical tool to adequately distribute the economic resources for maintenance and conservation for mid-term budget planning. In Chapter 6, and in the various appendices, data and results are presented of the developed application for the analyzed local network, from the optimization model, which guarantees the correct allocation of scarce resources at the disposal of authorities responsible for the regional networks in Spain. The implementation of these systems is witnessed with particular interest for the Spanish secondary network, because of the increasing management responsibility, with decreasing resources. Chapter 7 presents a series of conclusions that triggers to reconsider shifting from theoretical studies and conferences towards a practical implementation, considering how to properly conceive the engineering input and the related education. The original contributions of the document are also listed. In the same way, the research on the Bridges Management System can help evaluating and improving the used systematics.

Fluorescence correlation spectroscopy reveals fast optical excitation-driven intramolecular dynamics of yellow fluorescent proteins

Fast excitation-driven fluctuations in the fluorescence emission of yellow-shifted green fluorescent protein mutants T203Y and T203F, with S65G/S72A, are discovered in the 10−6–10−3-s time range, by using fluorescence correlation spectroscopy at 10−8 M. This intensity-dependent flickering is conspicuous at high pH, with rate constants independent of pH and viscosity with a minor temperature effect. The mean flicker rate increases linearly with excitation intensity for at least three decades, but the mean dark fraction of the molecules undergoing these dynamics is independent of illumination intensity over ≈6 × 102 to 5 × 106 W/cm2. These results suggest that optical excitation establishes an equilibration between two molecular states of different spectroscopic properties that are coupled only via the excited state as a gateway. This reversible excitation-driven transition has a quantum efficiency of ≈10−3. Dynamics of external protonation, reversibly quenching the fluorescence, are also observed at low pH in the 10- to 100-μs time range. The independence of these two bright–dark flicker processes implies the existence of at least two separate dark states of these green fluorescent protein mutants. Time-resolved fluorescence measurements reveal a single exponential decay of the excited state population with 3.8-ns lifetime, after 500-nm excitation, that is pH independent. Our fluorescence correlation spectroscopy results are discussed in terms of recent theoretical studies that invoke isomerization of the chromophore as a nonradiative channel of the excited state relaxation.

Multiple pathways for ultrafast transduction of light energy in the photosynthetic reaction center of Rhodobacter sphaeroides

A pathway of electron transfer is described that operates in the wild-type reaction center (RC) of the photosynthetic bacterium Rhodobacter sphaeroides. The pathway does not involve the excited state of the special pair dimer of bacteriochlorophylls (P*), but instead is driven by the excited state of the monomeric bacteriochlorophyll (BA*) present in the active branch of pigments along which electron transfer occurs. Pump-probe experiments were performed at 77 K on membrane-bound RCs by using different excitation wavelengths, to investigate the formation of the charge separated state P+HA−. In experiments in which P or BA was selectively excited at 880 nm or 796 nm, respectively, the formation of P+HA− was associated with similar time constants of 1.5 ps and 1.7 ps. However, the spectral changes associated with the two time constants are very different. Global analysis of the transient spectra shows that a mixture of P+BA− and P* is formed in parallel from BA* on a subpicosecond time scale. In contrast, excitation of the inactive branch monomeric bacteriochlorophyll (BB) and the high exciton component of P (P+) resulted in electron transfer only after relaxation to P*. The multiple pathways for primary electron transfer in the bacterial RC are discussed with regard to the mechanism of charge separation in the RC of photosystem II from higher plants.

On the absorbance changes in the photocycle of the photoactive yellow protein: A quantum-chemical analysis

Spectral changes in the photocycle of the photoactive yellow protein (PYP) are investigated by using ab initio multiconfigurational second-order perturbation theory at the available structures experimentally determined. Using the dark ground-state crystal structure [Genick, U. K., Soltis, S. M., Kuhn, P., Canestrelli, I. L. & Getzoff, E. D. (1998) Nature (London) 392, 206–209], the ππ* transition to the lowest excited state is related to the typical blue-light absorption observed at 446 nm. The different nature of the second excited state (nπ*) is consistent with the alternative route detected at 395-nm excitation. The results suggest the low-temperature photoproduct PYPHL as the most plausible candidate for the assignment of the cryogenically trapped early intermediate (Genick et al.). We cannot establish, however, a successful correspondence between the theoretical spectrum for the nanosecond time-resolved x-ray structure [Perman, B., Šrajer, V., Ren, Z., Teng, T., Pradervand, C., et al. (1998) Science 279, 1946–1950] and any of the spectroscopic photoproducts known up to date. It is fully confirmed that the colorless light-activated intermediate recorded by millisecond time-resolved crystallography [Genick, U. K., Borgstahl, G. E. O., Ng, K., Ren, Z., Pradervand, C., et al. (1997) Science 275, 1471–1475] is protonated, nicely matching the spectroscopic features of the photoproduct PYPM. The overall contribution demonstrates that a combined analysis of high-level theoretical results and experimental data can be of great value to perform assignments of detected intermediates in a photocycle.

Photogenerated o-Azaxylylenes: Mechanistic Studies and Synthetic Applications

This research sets out to build upon excited state o-azaxylylene cycloaddition. The mechanism behind the excitation and cycloaddition process of photogenerated o-azaxylylenes was determined experimentally. Time-correlated single-photon counting, steady-state spectroscopy, triplet quenching experiments, and quantum yield studies provided evidence suggesting that excited state intramolecular proton transfer is followed by intersystem crossing and stepwise addition to the tethered unsaturated pendant. In keeping with the principles of diversity oriented synthesis, a modular approach was taken to gain access to a diverse array of N,O,S-Polyheterocycles which were modified postphotochemically via Suzuki coupling to yield fused biaryls. Cycloaddition products, outfitted with halogens in the aromatic ring of the o-azaxylylene, proved to be reactive with a variety of boronic acids resulting in a rapid growth in structural complexity. A novel procedure was developed that utilized multiple o-azaxylylene cores in a photochemical cascade transformation yielding complex scaffolds of unprecedented topology. The photoprecursors were produced in a one-pot two-step sequence from commercially available starting materials, and upon irradiation yield structures containing up to five fused hetrocyclic rings, and showed complete diastereoselectivity.